Fig. 6

MFG-E8 increases phosphorylation and nuclear translocation of NF-κB p65 in aged VSMCs. a–h After being incubated with rMFG-E8 (a–b), Ang II (1 μM) (c–d), TNF-α (20 ng/mL) (e–f), and PBS (g–h) for 24 h, VSMCs isolated from young and aged aortas were immunostained with antibody against phosphorylated NF-κB p65 (Ser536). Arrows indicate nuclear translocation of NF-κB p65 in cells. Three independent experiments were performed; each experiment was repeated with similar results. i–j VSMCs isolated from the aortas of aged mice were cultured, either with or without rMFG-E8 (250 ng/mL), for 24 h. (i) Cell lysates were analyzed with immunoblotting with antibodies specific for NF-κB p65 phosphorylated at Ser536 and NF-κB p65. (j) Levels of phosphorylated p65, normalized to that of total p65 (n = 4), were analyzed. Data are presented as mean ± standard deviation. ****P < 0.0001; Student’s t test